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WO2025081564A1 - Souche de bifidobacterium breve permettant de soulager une déficience cognitive liée au vieillissement et utilisation associée - Google Patents

Souche de bifidobacterium breve permettant de soulager une déficience cognitive liée au vieillissement et utilisation associée Download PDF

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WO2025081564A1
WO2025081564A1 PCT/CN2023/131908 CN2023131908W WO2025081564A1 WO 2025081564 A1 WO2025081564 A1 WO 2025081564A1 CN 2023131908 W CN2023131908 W CN 2023131908W WO 2025081564 A1 WO2025081564 A1 WO 2025081564A1
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bbr16
bifidobacterium breve
probiotic
aging
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方曙光
盖忠辉
邵雅纹
董瑶
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WeCare Probiotics Co Ltd
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WeCare Probiotics Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745—Bifidobacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Definitions

  • the present application belongs to the technical field of probiotics, and relates to a Bifidobacterium breve BBr16 for improving cognitive impairment associated with aging and its application, and specifically to a Bifidobacterium breve BBr16 strain, a culture containing the same, a probiotic containing the same, and its application in the preparation of a medicine or health product for preventing, improving or treating cognitive impairment associated with aging.
  • the intestine contains approximately 1014 species of microorganisms, and the intestinal flora maintains a balance under normal physiological conditions.
  • the pathogenesis of cognitive impairment associated with aging is closely related to the imbalance of the gut-liver axis and enteric endotoxemia.
  • intestinal lipopolysaccharide can also promote inflammatory responses.
  • Bifidobacterium is a probiotic that is widely present in the human intestine and is considered to be an important component of the intestinal microecological balance. Bifidobacterium has good adhesion, can maintain the structure of the intestinal flora, and effectively protect the integrity of the intestinal mucosal morphology and metabolic function. Bifidobacterium plays an important role in the prevention and treatment of intestinal flora disorders. Its main mechanism includes regulating the balance of the flora through steric hindrance. In addition, bifidobacteria can also produce a variety of antibacterial components, such as acidic substances, antimicrobial peptides and biological enzymes, thereby inhibiting the growth and activity of pathogens. As a probiotic, bifidobacterium microecological preparations are safe, healthy, and have no side effects. They can colonize in animals and exert a variety of beneficial health effects. Therefore, they have been widely used in the field of clinical supplements.
  • anti-inflammatory cytokines According to the pathogenesis and clinical pathways of aging-related cognitive impairment, anti-inflammatory cytokines, antioxidants and methods of regulating intestinal flora can be selected to treat aging-related cognitive impairment. In this context, it is particularly important to develop more probiotic products to improve aging-related cognitive impairment.
  • the present application provides a method for improving aging-related cognitive impairment using Bifidobacterium breve BBr16 and its application
  • the invention specifically provides a Bifidobacterium breve BBr16 strain, a culture containing the same, a probiotic containing the same, and use of the same in preparing a medicine or health product for preventing, improving or treating cognitive impairment associated with aging.
  • the present application provides a Bifidobacterium breve for improving aging-related cognitive impairment.
  • the Bifidobacterium breve for improving aging-related cognitive impairment is named Bifidobacterium breve BBr16 strain, with a preservation number of CGMCC No. 24471 and a preservation date of March 7, 2022.
  • the present application isolates and preserves a new strain of Bifidobacterium breve that can prevent or treat cognitive impairment associated with aging from the feces of healthy breastfed infants in Bama Yao Autonomous County, Guangxi, and names it Bifidobacterium breve BBr16.
  • the strain can significantly improve cognitive impairment associated with aging, specifically: (1) significantly reducing the aging score of SAMP mice (rapidly aging mice); (2) significantly increasing the moving distance and residence time in the central area of the SAMP mice in a mine, and reducing the delay time to reach the platform in a maze experiment; (3) significantly increasing the dopamine and serotonin levels in the striatum and hippocampus of the SAMP mice; (4) reducing the levels of serum pro-inflammatory cytokine TNF- ⁇ and IL-6 in SAMP mice, and increasing the level of serum anti-inflammatory cytokine IL10; and (5) significantly increasing the SOD enzyme activity in the mouse liver, increasing the liver GSH content, increasing the liver CAT enzyme level, and reducing the liver MDA level. Therefore, the Bifidobacterium breve BBr16 strain can be used to prepare products for preventing, improving or treating aging-related cognitive impairment. As a probiotic, the BBr16 strain is highly safe and not prone to drug resistance.
  • the present application provides a culture of Bifidobacterium breve for improving aging-related cognitive impairment, wherein the preparation method of the culture comprises: inoculating the Bifidobacterium breve BBr16 strain described in the first aspect into a culture medium at 30-40°C (e.g., 30°C, 32°C, 35°C, 36°C, The cells are cultured at 37° C., 38° C., etc. for 12-30 h (e.g., 12 h, 14 h, 18 h, 20 h, 21 h, 22 h, 23 h, 24 h, 30 h, etc.).
  • 12-30 h e.g., 12 h, 14 h, 18 h, 20 h, 21 h, 22 h, 23 h, 24 h, 30 h, etc.
  • the culture medium may be MRS culture medium, and its formula composition exemplarily includes: peptone, beef extract, glucose, sodium acetate, yeast powder, diammonium hydrogen citrate, K 2 PO 4 ⁇ 3H 2 O, MgSO 4 ⁇ 7H 2 O, MnSO 4 , L-cysteine, and polysorbate 80 (Tween 80).
  • the present application provides a probiotic for preventing, improving or treating cognitive impairment associated with aging, wherein the strain in the probiotic includes the Bifidobacterium breve BBr16 strain described in the first aspect.
  • the Bifidobacterium breve BBr16 strain involved in the present application can be used alone in relevant products or in combination with other strains in relevant products.
  • the viable count of the Bifidobacterium breve BBr16 strain is not less than 1 ⁇ 10 8 CFU/mL or 1 ⁇ 10 8 CFU/g, for example, 1 ⁇ 10 8 CFU/mL (CFU/g), 1 ⁇ 10 9 CFU/mL (CFU/g), 2 ⁇ 10 9 CFU/mL (CFU/g), 5 ⁇ 10 9 CFU/mL (CFU/g), 8 ⁇ 10 9 CFU/mL (CFU/g), 1 ⁇ 10 10 CFU/mL (CFU/g), 5 ⁇ 10 10 CFU/mL (CFU/g) or 1 ⁇ 10 11 CFU/mL (CFU/g), etc.
  • Other specific point values within the numerical range can be selected, and they will not be described one by one here.
  • the dosage form of the probiotic includes lyophilized powder, capsule, tablet or granule.
  • the probiotic further comprises a protective agent and/or an auxiliary additive.
  • the protective agent comprises skimmed milk powder.
  • the auxiliary additives include any one of inulin, oligofructose, oligogalactose, oligomannose, trehalose, soybean oligosaccharide, resistant dextrin, spirulina, polydextrose, ⁇ -lactalbumin or lactoferrin, or a combination of at least two thereof.
  • the strains in the probiotic agent also include Lactobacillus paracasei LC86 strain; the preservation number of the Lactobacillus paracasei LC86 strain is CGMCC No.1.12731, and the preservation date is July 20, 2020.
  • the present application also creatively discovered that the above-mentioned BBr16 strain can be used in combination with the LC86 strain to prevent, improve or treat cognitive impairment associated with aging, and has significantly better effects than a single bacterial agent or other combination methods, indicating that the BBr16 strain and the LC86 strain have a synergistic effect in the above-mentioned effects.
  • the ratio of the viable cell counts of the Bifidobacterium breve BBr16 strain to the Lactobacillus paracasei LC86 strain is (3-5):1, for example, 3:1, 7:2, 4:1, 9:2 or 5:1, etc. Other specific point values within this numerical range can be selected, and they will not be described one by one here.
  • the two strains Based on the potential interaction between the BBr16 strain and the LC86 strain in the above-mentioned efficacy, the two strains have a better synergistic effect when the above-mentioned specific mass ratio relationship is met.
  • the present application provides the use of the Bifidobacterium breve BBr16 strain described in the first aspect, the culture described in the second aspect, or the probiotic agent described in the third aspect in the preparation of a medicine or health product for preventing, improving or treating cognitive impairment associated with aging.
  • the present application isolates and preserves a new strain of Bifidobacterium breve that can prevent or treat cognitive impairment associated with aging from fecal samples of healthy breastfed infants in Bama Yao Autonomous County, Guangxi, and names it Bifidobacterium breve BBr16.
  • the strain can significantly improve cognitive impairment associated with aging, specifically: (1) significantly reducing the aging score of SAMP mice (rapidly aging mice); (2) significantly increasing the moving distance and residence time in the central area of the SAMP mice in a mine field, and reducing the delay time to reach the platform in a maze experiment; (3) significantly increasing the dopamine and serotonin levels in the striatum and hippocampus of the SAMP mice; (4) reducing the levels of serum pro-inflammatory cytokine TNF- ⁇ and IL-6 in SAMP mice, and increasing the level of serum anti-inflammatory cytokine IL10; and (5) significantly increasing the SOD enzyme activity in the mouse liver, increasing the liver GSH content, increasing the liver CAT enzyme level, and reducing the liver MDA level. Therefore, the Bifidobacterium breve BBr16 strain can be used to prepare products for preventing, improving or treating aging-related cognitive impairment. As a probiotic, the BBr16 strain is highly safe and not prone to drug resistance.
  • FIG. 1 is a statistical diagram of the aging degree assessment results of each group of mice.
  • FIG2 shows the moving distance (cm) of mice in each group in the open field test.
  • FIG3 shows the time (s) that mice in each group stayed in the central area in the open field test.
  • FIG. 4 shows the delay time (s) for each group of mice to reach the platform in the water maze test.
  • FIG5 is a statistical graph showing the levels of dopamine and 5-hydroxytryptamine in the bilateral striatum and hippocampus of mice in each group.
  • Figure 6 shows the serum levels of tumor necrosis factor TNF ⁇ , interleukin 6 (IL6), MCP1 and IL10 in each group of mice Horizontal statistics chart.
  • FIG. 7 is a statistical graph showing the levels of superoxide dismutase (SOD), glutathione (GSH), peroxidase (CAT) and malondialdehyde (MDA) in the colon tissues of mice in each group.
  • SOD superoxide dismutase
  • GSH glutathione
  • CAT peroxidase
  • MDA malondialdehyde
  • the Bifidobacterium breve BBr16 strain involved below is deposited in the General Microbiology Center of China Culture Collection Administration, with the deposit number CGMCC No.24471, the deposit date is March 7, 2022, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.
  • Lactobacillus paracasei LC86 strain is deposited in the General Microbiology Center of China Culture Collection of Microorganisms, with the deposit number CGMCC No.1.12731. The deposit date is July 20, 2020, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.
  • MRS solid culture medium weigh 10 g of peptone, 10 g of beef extract, 20 g of glucose, 2 g of sodium acetate, 5 g of yeast powder, 2 g of diammonium hydrogen citrate, 2.6 g of K 2 PO 4 ⁇ 3H 2 O, 0.1 g of MgSO 4 ⁇ 7H 2 O, 0.05 g of MnSO 4 , 20 g of agar, and 0.5 g of L-cysteine, dissolve them in deionized water, add 1 mL of Tween 80, make up to 1 L, sterilize and cool, and pour into a sterilized culture dish for later use.
  • MRS liquid culture medium 10 g of peptone, 10 g of beef extract, 20 g of glucose, 2 g of sodium acetate, 5 g of yeast powder, 2 g of diammonium hydrogen citrate, 2.6 g of K 2 PO 4 ⁇ 3H 2 O, 0.1 g of MgSO 4 ⁇ 7H 2 O, 0.05 g of MnSO 4 and 0.5 g of L-cysteine were weighed, dissolved in deionized water, and 1 mL of Tween 80 was added to make up to 1 L, and the mixture was sterilized and cooled for later use.
  • the following method for preparing the bacterial suspension involves: inoculating the desired strain into MRS liquid culture medium, culturing at 37°C for 24 hours for activation, and continuously activating twice to obtain an activated solution; inoculating the activated solution into MRS liquid culture medium at an inoculum amount of 2% (v/v), culturing at 37°C for 24 hours to obtain a bacterial solution; centrifuging the bacterial solution at 8000g for 10 minutes, and resuspending the bacteria in PBS to obtain the obtained product.
  • the experimental animals were SPF-grade male SAMP8 mice, and mouse feed was purchased from Shanghai Slake Company; ELISA kits purchased from Shanghai ELISA Biotechnology Co., Ltd. were used to measure the levels of TNF ⁇ , IL1b, IL6 and MCP1; and detection kits purchased from Nanjing Jiancheng Bioengineering Company were used to measure the levels of superoxide dismutase (SOD), glutathione (GSH), peroxidase (CAT) and malondialdehyde (MDA).
  • SOD superoxide dismutase
  • GSH glutathione
  • CAT peroxidase
  • MDA malondialdehyde
  • a strain of Bifidobacterium breve that improves cognitive impairment associated with aging is screened, and the steps are as follows:
  • the pH of the MRS medium was adjusted to 3.0, and after sterilization at 121°C for 15 min, the liquid culture propagation material that had been activated for two generations was inoculated at a 2% inoculation rate, and cultured at 37°C for 24 h, and the change in absorbance ⁇ OD 600 value during 24 h was determined; 0.3% ox bile salt was added to the MRS medium, and after sterilization at 121°C for 15 min, the liquid culture that had been activated for two generations was inoculated at a 2% inoculation rate, and cultured at 37°C for 24 h, and the change in absorbance ⁇ OD 600 value during 24 h was determined, and finally 10 strains with two relatively large ⁇ OD 600 values were selected for the next experiment.
  • Bile salt tolerance experiment The liquid culture of the strain after two generations of activation was inoculated with 2% inoculum into MRS medium containing different bile salt concentrations (0.1%, 0.2%, 0.3%, 0.5%, and 2% bile salts in the culture medium, respectively). At the same time, MRS medium without bile salts was used as a control. After constant temperature culture at 37°C for 24 hours, samples were taken to determine the number of viable bacteria. Combined with the results of the previous experiment, excellent strains that are resistant to acid and bile salts were screened out.
  • the acid production capacity of the strain was determined by titration.
  • the strains preserved in glycerol tubes were activated and inoculated into MRS liquid medium at a 2% inoculum, and cultured at 37°C for 18 hours. 10 mL of the fermentation broth of each strain was added to 50 mL of sterile water, and 2-3 drops of 1 g/L phenolphthalein were added as an indicator. The solution was titrated with a 0.1 mol/L NaOH standard solution. The titration endpoint was when the solution turned pink and did not fade after 30 seconds. Each sample was repeated 3 times in parallel.
  • Example 1 the strains screened in Example 1 were subjected to physiological and biochemical characteristics and 16S rRNA molecular biological identification, and the steps were as follows:
  • the strain stored at -80°C was taken out and inoculated into a centrifuge tube containing 20mL MRS liquid medium at a ratio of 2%. After culturing at 37°C for 24h, it was centrifuged at 8000rpm for 15min, the supernatant was removed, and the bacteria were collected. The genome of the strain was extracted, and PCR amplification was performed by adding bacterial universal primers. The amplified product was sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing and identification. The 16S rDNA sequence of the strain was analyzed and shown as SEQ ID No: 1. The sequence obtained by sequencing was compared with the nucleic acid sequence in GeneBank, and the result showed that the strain was Bifidobacterium breve.
  • Example 2 Based on the results of 16S rDNA molecular biological identification and morphological identification in Example 2, it was confirmed that the strain belonged to Bifidobacterium breve and was named Bifidobacterium breve BBr16 strain.
  • the Bifidobacterium breve BBr16 strain was inoculated into MRS liquid culture medium and cultured at different temperatures of 10-50°C for 48 hours. During the culture process, the OD600 value of the culture solution was measured by an ELISA instrument at intervals. The results are shown in Table 2.
  • the artificial gastric juice contains 0.20% NaCl and 0.30% pepsin by mass fraction, and the pH is adjusted to 2.0, 2.5 and 3.0 respectively with HCl, and filtered for sterilization.
  • 10% ox bile salt weigh 10.0 g ox bile salt (Wokai, Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) into 100 mL sterile water and filter through a 0.22 ⁇ m filter membrane for sterilization.
  • MRS medium containing 0.1% bile salts MRS medium (Beijing Solebow Technology Co., Ltd.) was prepared according to the instructions. After high-pressure steam sterilization (121°C, 15 min), an appropriate amount of 10% ox bile salt solution was added to a final concentration of 0.1%.
  • N1 is the number of viable bacteria after 3 hours of artificial gastric juice treatment
  • N0 is the number of viable bacteria at 0 hours.
  • N1 is the number of viable bacteria after 3 hours of bile salt treatment
  • N0 is the number of viable bacteria at 0 hours. The test result shows that the survival rate is 32%.
  • the Bifidobacterium breve BBr16 described in the present application has good gastric acid resistance and choline resistance.
  • the good gastric acid and bile salt tolerance creates conditions for its colonization in the gastrointestinal tract, maintaining the homeostasis of the gastrointestinal mucosal barrier, and preparing products for preventing, improving or treating aging-related cognitive impairment.
  • mice Fifty 16-week-old adult male SAMP8 mice (22 ⁇ 2 g) were randomly divided into There were 5 groups, 10 mice in each group, including model control group (CTL group), Bifidobacterium breve BBr16 intervention group (BBr16 group), Lactobacillus paracasei LC86 intervention group (LC86 group), Bifidobacterium breve BBr16 and Lactobacillus paracasei LC86 composite bacteria intervention group (BBr16+LC86 group, viable bacteria ratio 4:1), and Bifidobacterium breve BBr16 and Lactobacillus paracasei ATCC27092 composite bacteria intervention group (BBr16+ATCC27092 group, viable bacteria ratio 4:1).
  • CTL group model control group
  • BBr16 group Bifidobacterium breve BBr16 intervention group
  • LC86 group Lactobacillus paracasei LC86 intervention group
  • BBr16+LC86 group, viable bacteria ratio 4:1 Bifidobacterium breve BB
  • mice 10 16-week-old adult male SAMP8 mice (22 ⁇ 2g) were used as young control group (Young group). All mice were free of specific pathogens. The mice were raised individually in cages in a clean and quiet environment with a temperature of 23-25°C and a humidity of 50-70%. Mice in each group had free access to food and water during the experimental period.
  • mice in the above groups were evaluated using a grading system. Each category has 5 levels, recorded as 0, 1, 2, 3, and 4, respectively. The higher the score, the more severe the aging.
  • the indicators used in the evaluation include reactive behavior, passive behavior, gloss, roughness, hair loss, skin ulcers, periocular lesions, and spinal lordosis. The scores of each aspect were added together to obtain the score of each mouse. The scoring criteria for mouse aging are shown in Table 4.
  • Open field test method The mice were gently transferred to a plexiglass activity chamber of 25 ⁇ 25 ⁇ 40 cm in size. The mice were allowed to freely explore the activity chamber for 10 minutes. The behavioral performance of the mice in the open field activity chamber was observed and recorded, including the moving distance (cm) and the residence time (s) in the central area. After each experiment, the chamber was cleaned with 70% ethanol to reduce odor interference. The statistical results are shown in Figures 2 and 3 (presented as average values).
  • the results in the figure above show that BBr16 can significantly improve cognitive impairment caused by aging in mice.
  • the BBr16+LC86 group had the best improvement ability, which was higher than the BBr16 group and the LC86 group, indicating that the BBr16 strain and the LC86 strain have a synergistic effect in improving cognitive impairment caused by aging in mice.
  • the samples were homogenized and centrifuged at 12000g and 4°C for 10 minutes. The supernatant was filtered using a 0.22 ⁇ m membrane, and the filtrate was used for HPLC-MS detection.
  • HPLC part the Thermo Vanquish ultra-high performance liquid phase system of Thermo Fisher Scientific was used, and the ACQUITY UPLC HSST3 column (2.1 ⁇ 150mm, 1.8 ⁇ m) of Waters was used. The flow rate was set to 0.25mL/min, the column temperature was 40°C, and the injection volume was 2 ⁇ L.
  • BBr16 can significantly increase the levels of dopamine and 5-HT in the bilateral striatum and hippocampus of SAMP8 mice.
  • the BBr16+LC86 group had the largest increase, which was better than the BBr16 group and the LC86 group.
  • the BBr16 strain has an excellent effect in improving aging-related cognitive impairment, and the BBr16 strain and the LC86 strain have a synergistic effect in the above-mentioned effects.
  • Enzyme-linked immunosorbent assay was used to detect the levels of serum tumor necrosis factor TNF ⁇ , interleukin 6 (IL6), MCP1, and IL10 in each group of mice.
  • Serum samples were collected from each group, and the levels of serum tumor necrosis factor TNF ⁇ , interleukin 6 (IL6), MCP1 and IL10 were determined using ELISA kits. The results are shown in Figure 6. It can be seen from the results that BBr16 intervention can reduce the production of serum TNF ⁇ , IL6 and MCP1, and promote the production of serum IL10, and the effect of the BBr16+LC86 group is more obvious.
  • mice were killed, and the colons of the mice in each group were added to normal saline and then homogenized to obtain 10% colon homogenate.
  • the homogenate was centrifuged for 10 min (5,000 g, 4°C) to obtain the supernatant.
  • SOD superoxide dismutase
  • GSH glutathione
  • CAT peroxidase
  • MDA malondialdehyde
  • the present application uses the above embodiments to illustrate a method of improving aging-related Bifidobacterium breve for cognitive impairment and its application, but the present application is not limited to the above embodiments, that is, it does not mean that the present application must rely on the above embodiments to be implemented.
  • the technicians in the relevant technical field should understand that any improvement of the present application, the equivalent replacement of the raw materials of the product of the present application, the addition of auxiliary components, the selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present application.

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Abstract

L'invention concerne une souche de Bifidobacterium breve permettant de soulager une déficience cognitive liée au vieillissement et une utilisation associée. Ladite souche est nommée Bifidobacterium breve BBr16 et le numéro d'accès associé est CGMCC No. 24471 La souche peut soulager remarquablement une déficience cognitive liée au vieillissement et peut être utilisée pour préparer des produits ayant des effets de prévention, de soulagement ou de traitement d'une déficience cognitive liée au vieillissement.
PCT/CN2023/131908 2023-10-16 2023-11-16 Souche de bifidobacterium breve permettant de soulager une déficience cognitive liée au vieillissement et utilisation associée Pending WO2025081564A1 (fr)

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